The purpose of this non-clinical, IIDEA project is to study the feasibility of using retroviral vectors to transfer the gene which is abnormal in autosomal recessive Chronic Granulomatous Disease(p47-phox) into human hematopoietic cells. Development of retrovirus producer cell lines capable of infecting human blood cells will provide the opportunity to study expression of this protein in precursor and differentiated hematopoietic cells. A cDNA for p47-phox containing the protein coding-sequence has been cloned into a retroviral vector known as pLXSN. Two murine producer cell lines have been established, one producing an ecotropic retrovirus capable of infecting rodent cells and the other producing amphotropic retrovirus capable of infecting a wide range of mammalian and avian cells. The amphotropic virus containing p47-phox in both normal(sense) and reverse(antisense) orientations was used to infect two leukemia cell lines, HL-60 and U937. RNA and protein analysis demonstrated retrovirally transferred p47-phox genes in these cells. In addition, Epstein Barr Virus transformed B lymphocytes from a normal individual and an AR-CGD patient were also infected. These lines also appear to have been transduced by the retroviruses based on protein and RNA analysis.Assays to determine whether the protein made from the transferred gene is functional are still in progress. EBV transformed B lymphocyte cell lines derived from AR-CGD patient cells may be helpful in studies of correction of the defect since these cells in normals produce a small amount of superoxide and can be studied at the functional as well as molecular level. Expression of transferred p47-phox in murine bone marrow and hematopoietic reconstitution of an irradiated mouse will be necessary before primate and human studies can proceed. Preliminary experiments indicate that it is possible to transfer this gene into murine bone marrow.